Culture and amplification-free bacterial sepsis diagnosis

无培养和扩增细菌败血症诊断

• 批准号: 10805776
• 负责人: Alfredo Andres Celedon
• 金额: $ 100万
• 依托单位: SCANOGEN, INC.
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-09 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10805776
• 关键词: Culture; amplification; free; bacterial; sepsis

Culture and amplification-free bacterial sepsis diagnosis Summary Sepsis is a life-threatening complication of an infection that requires immediate treatment. The identification of the causative microorganism utilizing processes based on blood culture is a keystone of sepsis diagnosis. However, blood culture takes one to five days to provide results. Here, we propose the development of a culture and amplification-free assay to detect and identify bacteria in one hour directly on whole blood. The new assay will enable physicians to provide therapy targeting the causative organism days earlier than the current standards. The new assay will be based on Single MOLecule Tethering (SMOLT), a novel sample preparation and amplification-free molecular detection method developed at Scanogen. The work proposed in this application will build on the achievements of an ongoing project aimed at developing an assay to detect fungi in whole blood. The SMOLT-Bacteria assay proposed in this application will identify the species, genus or order of the causative organism in approximately 90% of bloodstream infections. The new system will be easy-to-use, fully automated, and capable of rapid detection in whole blood without blood culture. Despite the urgent clinical need, there is only one FDA approved culture-free molecular assay for sepsis diagnosis. In comparison to this test, the SMOLT- Bacteria assay will be 4 times faster, 6 times less expensive, and thanks to its higher multiplexing capability, it will detect and identify the causative pathogen in twice the number of patients. In our preliminary results, we developed a multiplex assay capable of detecting bacteria directly in whole blood. We demonstrated that the assay is highly specific and sensitive with a limit of detection (LOD) between 1-10 CFU/mL, depending on the species, which is as good as the only FDA-approved test. In this project, we will complete the development of the assay protocol by including new probes and optimizing the overall assay (Aim 1). We will test the manual protocol and develop a fully integrated disposable cartridge and instrument (Aim 2). Finally, we will evaluate the assay in pilot analytical and clinical validation studies (Aim 3). We will work with a multidisciplinary team that includes experts in assay development and instrument development, including the former Vice President of Engineering at Becton Dickinson, experts in microbiology, sepsis diagnosis and treatment from Johns Hopkins University and the University of Pittsburgh, as well as engineers from Key Technologies. After completing this project, we will further optimize the system for usability and manufacturability and conduct analytical and clinical studies for submission to the Food and Drug Administration (FDA). If successful, the new automated assay can become part of the standard procedure for the diagnosis of patients with sepsis and improve patients’ outcomes by dramatically reducing the time to targeted antimicrobial therapy.

细菌败血症的培养和无扩增诊断 摘要 脓毒症是一种威胁生命的感染并发症，需要立即治疗。身份的鉴定 基于血培养的病原微生物利用过程是脓毒症诊断的重点。 然而，血液培养需要一到五天的时间才能产生结果。在这里，我们建议发展一种文化 以及无需扩增的检测方法，可在一小时内直接从全血中检测和鉴定细菌。新的化验方法 将使医生能够在比目前更早的几天提供针对致病微生物的治疗 标准。 新的分析将基于单分子连接(SMOLT)，这是一种新的样品制备和 Scanogen开发了无扩增的分子检测方法。本申请中提出的工作 将建立在一个正在进行的项目的成果基础上，该项目旨在开发一种检测全血中真菌的方法。 在本申请中提出的烟雾细菌检测将鉴定致病原的种类、属或目。 在大约90%的血液感染中存在细菌。新系统将易于使用，完全自动化， 且无需血培养即可快速检测全血。尽管临床上有迫切的需求，但有 只有一家FDA批准了用于脓毒症诊断的无培养分子检测。与这次测试相比，Smoolt- 细菌检测速度将提高4倍，成本降低6倍，而且由于其更高的多重能力，它 将在两倍数量的患者中检测和识别致病病原体。 在我们的初步结果中，我们开发了一种能够直接检测全血中细菌的多重检测方法。 我们证明了该方法具有高度的特异性和敏感性，检测限在1-10之间。 Cfu/ml，取决于物种，这与FDA批准的唯一测试一样好。在这个项目中，我们将 通过加入新的探针和优化整体检测(AIM)来完成检测方案的开发 1)。我们将测试手动协议，并开发完全集成的一次性墨盒和仪器(目标2)。 最后，我们将在试验性分析和临床验证研究(目标3)中对该分析进行评估。我们将与一个 多学科团队，包括分析开发和仪器开发方面的专家，包括 贝顿·迪金森公司前工程副总裁，微生物学、脓毒症诊断和治疗专家 来自约翰霍普金斯大学和匹兹堡大学的治疗，以及来自Key的工程师 技术。在完成这个项目后，我们将进一步优化系统的可用性和可制造性 并进行分析和临床研究，提交给食品和药物管理局(FDA)。如果 如果成功，新的自动化化验可以成为患者诊断的标准程序的一部分 治疗脓毒症，并通过显著减少靶向抗菌治疗的时间来改善患者的预后。